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大黄素对角膜基质细胞分泌细胞因子的干预作用

陈国玲1,2,陈双3,刘艳丽1,魏璐婉1,刘执玉1
  

  1. (1. 山东大学医学院人体解剖与组织胚胎学研究所, 济南 250012;
    2. 山东大学眼科中心, 第二医院眼科, 济南 250033;
    3. 枣庄市山亭区人民医院, 山东 枣庄 277200)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 发布日期:2009-04-16
  • 通讯作者: 刘执玉

Inhibitory effect of emodin on cytokine secrete from cultured corneal fibroblasts

CHEN Guoling1,2, CHEN Shuang3, LIU Yanli1, WEI Luwan1, LIU Zhiyu1
  

  1. (1. Department of Anatomy, School of Medicine, Shandong University, Jinan 250012, China;
    2. Ophthalmic Center, Department of Ophthalmology, Second Hospital of Shandong University, Jinan 250033, China;
    3. People′s Hospital of Shanting District, Zaozhuang 277200, Shandong, China)
  • Received:1900-01-01 Revised:1900-01-01 Published:2009-04-16
  • Contact: LIU Zhiyu

摘要: 目的观察大黄素对体外培养的人眼角膜基质细胞分泌白细胞介素6(IL6)的干预作用。方法四甲基偶氮唑蓝(MTT)法检测不同浓度大黄素对人眼角膜基质细胞活性的影响。绿脓杆菌脂多糖(LPS)刺激角膜基质细胞(LPS组),并用大黄素进行干预(大黄素预处理组)。分别于LPS刺激前(0?h),刺激后1、2、4、8?h收集各组细胞及细胞上清液。Western blot检测不同时点角膜基质细胞κB抑制因子α(IκBα)蛋白的表达及大黄素的作用,酶联免疫吸附实验(ELISA)检测不同时点角膜基质细胞IL6蛋白的分泌及大黄素的作用。结果不同浓度大黄素对角膜基质细胞的活性没有影响。与0?h相比,LPS刺激角膜基质细胞1?h时,胞浆IκBα蛋白表达出现下降,其电泳条带随着刺激时间的延长而逐渐变暗,4?h时最暗。LPS刺激后各时点IκBα蛋白表达较LPS刺激前均明显下降(P<0.01)。LPS刺激可引起人眼角膜基质细胞IL6蛋白分泌增多,刺激后8h达峰值,LPS刺激细胞后各时点细胞分泌IL6均较刺激前明显增加(P<0.01)。大黄素预处理后,与LPS组相比较,角膜基质细胞表达IκBα蛋白下降、细胞分泌IL6减少(P<0.01)。结论绿脓杆菌LPS可引起人眼角膜基质细胞IκBα降解,引起细胞因子表达,在角膜炎的过程中可能起一定的作用。大黄素能有效抑制LPS诱导的人角膜基质细胞IκBα降解,从而减少相应细胞因子的分泌。

关键词: 大黄素, 白细胞介素6, 角膜

Abstract:

MethodsCell viability insensitive to emodin on cultured human corneal fibroblasts was assessed using the MTT assay. Human corneal fibroblasts were randomly divided into three groups:the control group(group 0?h), the lipopolysaccharide(LPS) group and the emodin pretreatment group. Cells in the emodin pretreatment group were incubated with emodin for 30?min before being LPS challenged. Before and after 1,2,4 and 8?h treatment with LPS, expression of inhibitor of κBα(IκBα)was assayed by Western blot, and secretion of IL6 induced by LPS from cultured corneal fibroblasts was determined with enzymelinked immunosorbent assays(ELISA). Effects of emodin, an active component from the rhizome of Rheum palmatum, on expressions of IκBα and IL6 were also assessed in these cells. ResultsThe concentration of emodin used in this study was safe for cultured human corneal fibroblasts. Compared with the 0h group, expression of IκBα protein level was significantly decreased after being LPS challenged 18?h(P<0.01), the lowest expression of IκBα was observed at 4h after LPS challenged. Compared with cells in the 0h group, IκBα level was significantly decreased in each group after treated with LPS, and degeneration of IκBα induced by LPS was markedly inhibited by emodin(P<0.01). At the same time, the protein release of IL6 in corneal fibroblasts was significantly increased at each point after being challenged with LPS, which was significantly inhibited by pretreatment with emodin(P<0.01). ConclusionThese results suggest that LPS takes part in the degeneration of IκBα in cultured human corneal fibroblasts in vitro, and emodin can partly inhibit this degeneration. By inhibiting IκBα degeneration, emodin may suppress LPSinduced cytokine release from these cells, which is one of the signal transduction mechanisms of emodin to prevent the occurrence of inflammation.

Key words: Emodin, Interleukin6, Cornea 核因子

中图分类号: 

  • R772.2
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